Toolkit for evaluating genes required for proliferation and survival using tetracycline-regulated RNAi View Full Text


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Article Info

DATE

2011-01

AUTHORS

Johannes Zuber, Katherine McJunkin, Christof Fellmann, Lukas E Dow, Meredith J Taylor, Gregory J Hannon, Scott W Lowe

ABSTRACT

Short hairpin RNAs (shRNAs) are versatile tools for analyzing loss-of-function phenotypes in vitro and in vivo. However, their use for studying genes involved in proliferation and survival, which are potential therapeutic targets in cancer and other diseases, is confounded by the strong selective advantage of cells in which shRNA expression is inefficient. We therefore developed a toolkit that combines Tet-regulated miR30-shRNA technology, robust transactivator expression and two fluorescent reporters to track and isolate cells with potent target knockdown. We demonstrated that this system improves the study of essential genes and was sufficiently robust to eradicate aggressive cancer in mice by suppressing a single gene. Further, we applied this system for in vivo negative-selection screening with pooled shRNAs and propose a streamlined, inexpensive workflow that will facilitate the use of RNA interference (RNAi) for the identification and evaluation of essential therapeutic targets. More... »

PAGES

79

Identifiers

URI

http://scigraph.springernature.com/pub.10.1038/nbt.1720

DOI

http://dx.doi.org/10.1038/nbt.1720

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1005914891

PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/21131983


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JSON-LD is a popular format for linked data which is fully compatible with JSON.

curl -H 'Accept: application/ld+json' 'https://scigraph.springernature.com/pub.10.1038/nbt.1720'

N-Triples is a line-based linked data format ideal for batch operations.

curl -H 'Accept: application/n-triples' 'https://scigraph.springernature.com/pub.10.1038/nbt.1720'

Turtle is a human-readable linked data format.

curl -H 'Accept: text/turtle' 'https://scigraph.springernature.com/pub.10.1038/nbt.1720'

RDF/XML is a standard XML format for linked data.

curl -H 'Accept: application/rdf+xml' 'https://scigraph.springernature.com/pub.10.1038/nbt.1720'


 

This table displays all metadata directly associated to this object as RDF triples.

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